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Our aim was to evaluate whether repetition of C‐ion (carbon ion beam) irradiation induces radioresistance as well as repeated X‐ray irradiation in cancer cell lines, and to find the key molecular pathway for radioresistance by comparing radioresistant cancer cells with their parental cells. A mouse squamous cell carcinoma cell line, NR‐S1, and radioresistant cancer cells, NR‐S1‐C30 (C30) and NR‐S1‐X60 (X60), established by repetition of C‐ion and X‐ray irradiation, respectively, were used. X‐ray and C‐ion sensitivity, changes in lysosome, mitochondria, intracellular ATP and reactive oxygen species (ROS) level, and mechanistic target of rapamycin (mTOR) signaling were evaluated. Moreover, the effect of rapamycin on radioresistance was also assessed. X‐ray and C‐ion resistance of C30 cells was moderate, and the resistance of X60 cells was the highest in this study. In X60 cells, the amount of lysosome, mitochondria, intracellular ATP and ROS level were significantly increased, and mTOR and p70S6K (ribosomal protein S6 kinase p70) phosphorylation were enhanced compared with C30 and NR‐S1 cells. The inhibition of mTOR signaling was effective for X‐ray and C‐ion radiosensitization in both cell lines, especially in X60 cells in which X‐ray and C‐ion resistance was decreased to the same level as that in NR‐S1 cells. Our results indicated that the contribution to generate X‐ray and C‐ion resistance was less for repeated C‐ion irradiations compared with repeated X‐ray irradiation. Moreover, we found that activated mTOR signaling contributes to X‐ray and C‐ion resistance in the X60 cancer cells.

Enhancement of mTOR signaling contributes to acquired X‐ray and C‐ion resistance in mouse squamous carcinoma cell line